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KMID : 0545120190290030392
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Journal of Microbiology and Biotechnology
2019 Volume.29 No. 3 p.392 ~ p.400
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Biosynthesis of Chondroitin in Engineered Corynebacterium glutamicum
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Cheng Fangyu
Luozhong Sijin
Yu Huimin
Guo Zhigang
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Abstract
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Chondroitin, the precursor of chondroitin sulfate, which is an important polysaccharide, has drawn significant attention due to its applications in many fields. In the present study, a heterologous biosynthesis pathway of chondroitin was designed in a GRAS (generally recognized as safe) strain C. glutamicum. CgkfoC and CgkfoA genes with host codon preference were synthesized and driven by promoter Ptac, which was confirmed as a strong promoter via GFPuv reporter assessment. In a lactate dehydrogenase (ldh) deficient host, intracellular chondroitin titer increased from 0.25 to 0.88 g/l compared with that in a wild-type host. Moreover, precursor enhancement via overexpressing precursor synthesizing gene ugdA further improved chondroitin titers to 1.09 g/l. Chondroitin production reached 1.91 g/l with the engineered strain C. glutamicum ΔL-CgCAU in a 5-L fed-batch fermentation with a single distribution Mw of 186 kDa. This work provides an alternative, safe and novel means of producing chondroitin for industrial applications.
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KEYWORD
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Chondroitin biosynthesis, engineered Corynebacterium glutamicum, lactate dehydrogenase deficient, precursor enhancement, fed-batch fermentation
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